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Partial karyotypes and FISH images of genetic abnormalities associated with MM. (A and B) The partial karyotype shows the normal chromosome 14 and the normal chromosome 20 on the left of each pair. Additional material can be seen on the long arm of the derivative chromosome 14, from the derivative chromosome 20 depicting the t(14;20)(q32;q11). The FISH image using the Cytocell probe set for IGH and MAFB confirms that the rearrangement involves the juxtaposition of IGH and MAFB. (C) FISH image depicting the Vysis MYC break apart probe. There is an intact copy of the probe on one chromosome 8. The second fusion has broken apart but also demonstrates associated complexity with duplication of the centromeric part of the probe and involvement of three chromosomes. (D and E) The partial karyotype shows a normal chromosome 1 on the left with an isochromosome 1q on the right. The isochromosome shows two copies of the long arm mirror imaged around the centromere with effective loss of the short arm of chromosome 1. The FISH image shows the CytocellCDKN2C/CKS1B probe that highlights a gene of interest on the long arm in red and a gene of interest on the short arm of chromosome 1. A three red and one green signal pattern confirms the gain of 1q and the loss of 1p. Coloured arrows demonstrate the probe positions, green and red arrows show green and red signals, respectively; yellow arrows depict fusion signals.
Source publication
Despite advances in the treatment of multiple myeloma (MM), it remains an incurable malignant disease. Myeloma genetics is intrinsically complex, but it offers an opportunity to categorize the disease and apply a personalized medicine approach.
Research into the genetics of myeloma is moving at a fast pace and is highlighting areas and patient coho...
Contexts in source publication
Context 1
... be mediated by a fragile site in the WWOX gene on the long arm of chromosome 16. 17,23 This results in MAF coming under the regulatory influence of IGH, which in turn has the effect of up-regulating CCND2. 19 MAF rearrangements have been associated with a more aggressive clinical course. 23 Data on MAFB rearrange- ments associated with t(14;20) ( Fig. 1A and B) are not robust due to their rarity, although a similar clinical course would be predicted. 23 Translocations not involving IGH do occur, but they are considered unusual and are most likely to be seen in progressive disease. CCND3-MAF, MAF-FGFR3/MMSET, CCND3-FGFR3/MMSET re- arrangements have all been described. 19 MYC ...
Context 2
... disease. 21,23 MYC translo- cations are not considered to be initiating events, but rather late events associated with increased prolifer- ation and stromal independent plasma cells. 21 They are frequently seen as non-reciprocal translocation events involving more than one chromosome and associated regions of amplification and duplication (Fig. 1C). 21 The t(8;14)(q24;q32) accounts for only 25% of MYC rearrangement, 17,36 and recent studies have demonstrated that MYC is able to recruit active super enhancers from highly expressed genes asso- ciated with B cell, plasma cell or myeloma develop- ment. 37 Examples include enhancers associated with CCND1, XBP1, KRAS, FAM46C and ...
Context 3
... it can be present in different guises in different cells. Implicated genes include CKS1B and ANP32E. 21 Duplication of 1q is also associated with poor prognosis, although the intrinsic relationship with deletion 1p creates difficulties in assessing these abnormalities separately. 21 A common manifestation of del1p/dup1q is the isochromosome 1q, (Fig. 1D and E), where the short arm is lost and the long arm is duplicated and mirrored around the ...
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Multiple myeloma (MM) is a diverse clonal plasma cell malignancy with resultant organ damage including renal and bone marrow effects, as well as neurologic and immune dysfunction. MM is characterized as clinically and pathologically heterogeneous with significant variability in treatment response and survival. The genetically high-risk myeloma is o...
Treatment options in multiple myeloma (MM) are increasing with the introduction of complex multi-novel-agent-based regimens investigated in randomized clinical trials. However, application in the real-world setting, including feasibility of and adherence to these regimens, may be limited due to varying patient-, treatment-, and disease-related fact...
Multiple myeloma (MM) is an incurable malignancy of plasma cells that grow within a permissive bone marrow microenvironment (BMM). The bone marrow milieu supports the malignant transformation both by promoting uncontrolled proliferation and resistance to cell death in MM cells, and by hampering the immune response against the tumor clone. Hence, it...
Citations
... Indeed, conventional cytogenetics analysis assesses the presence of chromosomal abnormalities in metaphases of proliferating cells under standard culture conditions, while proliferation rate of plasma cells is limited and requires certain stimuli in culture medium, such as lipopolysaccharide, various cytokines (e.g., interleukin[IL]-6), or synthetic DSP30, an oligonucleotides containing CpG motif [38]. However, conventional karyotyping with plasma cell enrichment is not always feasible in routinely clinical practice and fails to recognize cryptic translocations of the IgH locus or 17p deletions [39]. Here, we documented a comparable sensitivity of NIPT to other methodologies in detecting chromosomal alterations in MM, even using peripheral blood specimens and without plasma cell enrichment procedures, especially in those subjects with extramedullary disease. ...
Liquid biopsy is a minimally invasive diagnostic tool for identification of tumor-related mutations in circulating cell-free DNA (cfDNA). The aim of this study was to investigate feasibility, sensitivity, and specificity of non-invasive prenatal test (NIPT) for identification of chromosomal abnormalities in cfDNA from a total of 77 consecutive patients with non-Hodgkin B-cell lymphomas, Hodgkin lymphoma (HL), or plasma cell dyscrasia. In this case series, half of patients had at least one alteration, more frequently in chromosome 6 (23.1%), chromosome 9 (20.5%), and chromosomes 3 and 18 (16.7%), with losses of chromosome 6 and gains of chromosome 7 negatively impacting on overall survival (OS), with a 5-year OS of 26.9% and a median OS of 14.6 months, respectively (P = 0.0009 and P = 0.0004). Moreover, B-cell lymphomas had the highest NIPT positivity, especially those with aggressive lymphomas, while patients with plasma cell dyscrasia with extramedullary disease had a higher NIPT positivity compared to conventional cytogenetics analysis and a worse outcome. Therefore, we proposed a NIPT-based liquid biopsy a complementary minimally invasive tool for chromosomal abnormality detection in hematological malignancies. However, prospective studies on larger cohorts are needed to validate clinical utility of NIPT-based liquid biopsy in routinely clinical practice.
... Although single nucleotide polymorphism (SNP) array has been used to identify copy number abnormalities, it does not identify fusions and the results have to be integrated with FISH analysis. 17,18 Transcriptomic analysis has been used to identify highrisk gene expression signatures. 19,20 This review will focus on FISH-based approach to identify the genetic subtypes. ...
... Hence, FISH is a superior tool with better sensitivity and specificity in identifying the clinically relevant genetic subtypes. 17,24 Fluorescent in-situ Hybridization FISH does not require live cells and can be performed on interphase cells. Plasma cell infiltration of the bone marrow can be patchy in MM, and the percentage of plasma cells in the marrow may vary. ...
Plasma cell dyscrasias are a heterogeneous group of neoplasms characterized by abnormal proliferation of plasma cells with or without over production of monoclonal immunoglobulins. Chromosomal abnormalities are acquired either early in the course of the disease or during disease progression. Plasma cell dyscrasias are categorized into multiple cytogenetic subtypes that form an integral component of risk-stratified treatment protocols. The primary genetic events are IgH gene translocations and non-random gains of chromosomes 3/5/7/9/11/15/19 and or 21. The secondary genetic events consist of chromosome 1 abnormalities (1p deletion and 1q gain or amplification), deletion 17p/TP53, deletion 13q, and MYC gene rearrangements. Plasma cells being at the end of differentiation spectrum of B cells, have low proliferative potential precluding the use of karyotyping in identification of chromosomal abnormalities. Analysis of enriched plasma cells using interphase fluorescent in situ hybridization (FISH) is the technique of choice for identifying these abnormalities. It is essential to enrich plasma cells before the FISH analysis, and numerous plasma cell enrichment techniques have been described. In the paper, we review the cytogenetic approach to identify clinically significant genetic aberrations including the effective use of FISH panels and plasma cell enrichment techniques.
... Multiple myeloma (MM) is derived from the development of clonal plasma cells (PCs) and the mass synthesis of monoclonal proteins in the bone marrow which eventually leads to end-organ failure [1]. The older population is the most affected by MM, with a median diagnosed age of 69 years [2], which is closely associated with a dismal prognosis and has a 5-year survival rate of 48.5% [3]. It is hypothesised that the MM clone originates from a post-germinal centre as a result of isotope switching and produces a PC that could continue to multiply endlessly [4]. ...
Multiple myeloma (MM) is an exceptionally complicated and heterogeneous disease that is caused by the abnormal proliferation of malignant monoclonal plasma cells initiated in the bone marrow. In disease progression, a multistep process including differentiation, proliferation, and invasion is involved. Despite great improvement in treatment outcomes in recent years due to the substantial discovery of novel therapeutic drugs, MM is still regarded as an incurable disease. Patients with MM are afflicted by confronting remission periods accompanied by relapse or progression outcomes, which inevitably progress to the refractory stage. In this regard, MM may need new medications or modifications in therapeutic strategies to overcome resistance. A variety of genetic abnormalities (e.g., point mutations, translocations, and deletions) and epigenetic changes (e.g., DNA methylation, histone modification, and non-coding RNA) contribute to the pathogenesis and development of MM. Here, we review the significant roles of epigenetic mechanisms in the development and progression of MM. We also highlight epigenetic pathways as potential novel treatment avenues for MM, including their interplay, use of epigenetic inhibitors, and major involvement in immuno-oncology.
... [2][3][4][5][6][7][8][9][10] Plasma cells accumulate in bone marrow (BM) leading to bone destruction and marrow failure. [10][11][12][13][14][15][16] Diagnosis is based on clinical, radiologic, and pathological characteristics. [7][8] For risk stratification, prognosis, and treatment efficacy, PCM is classified into high, standard, and low-risk clinical categories based on serum M-protein concentration, percent of plasma cells in the marrow (extent of bone marrow involvement), and identification of genetic abnormalities. ...
... [7][8] For risk stratification, prognosis, and treatment efficacy, PCM is classified into high, standard, and low-risk clinical categories based on serum M-protein concentration, percent of plasma cells in the marrow (extent of bone marrow involvement), and identification of genetic abnormalities. [9][10][11][12][13][14][15][16] The genetic abnormalities usually reflect various underlying pathways of clonal heterogeneity and subsequent evolution. [8][9][10][11] The genetic alterations are critical for prognosis, risk stratification, expected patient outcome, survival rate, and in selecting an appropriate therapeutic strategy. ...
... Deletion of chromosome 1p, gain of copy number or amplification of chromosome 1q, and abnormalities of TP53 locus have been reported to indicate PCM disease progression. [12][13][14][15][16][17] Genetic aberrations observed in standard to low-risk PCM include t (11;14), t (6;14), deletion or loss of chromosome 13, and hyperdiploidy. [10,[17][18][19][20] Deleted 13q is a negative prognostic indicator when observed in metaphase (dividing cells) cytogenetic analysis. ...
Background:
Plasma cell neoplasm and/or plasma cell myeloma (PCM) is a mature B-cell lymphoproliferative neoplasm of plasma cells that secrete a single homogeneous immunoglobulin called paraprotein or M-protein. Plasma cells accumulate in the bone marrow (BM) leading to bone destruction and BM failure. Diagnosis of PCM is based on clinical, radiologic, and pathological characteristics. The percent of plasma cells by manual differential (bone marrow morphology), the white blood cell (WBC) count, cytogenetics, fluorescence in situ hybridization (FISH), microarray, and next-generation sequencing of BM are used in the risk stratification of newly diagnosed PCM patients. The genetics of PCM is highly complex and heterogeneous with several genetic subtypes that have different clinical outcomes. National Comprehensive Cancer Network guidelines recommend targeted FISH analysis of plasma cells with specific DNA probes to detect genetic abnormalities for the staging of PCM (4.2021). Recognition of risk categories through training software for classification of high-risk PCM and a novel way of addressing the current approaches through bioinformatics will be a significant step toward automation of PCM analysis.
Methods:
A new artificial neural network (ANN) classification model was developed and tested in Python programming language with a first data set of 301 cases and a second data set of 176 cases for a total of 477 cases of PCM at diagnosis. Classification model was also developed with support vector machines (SVM) algorithm in R studio and interactive data visuals using Tableau.
Results:
The resulting ANN algorithm had 94% accuracy for the first and second data sets with a classification summary of precision (PPV): 0.97, recall (sensitivity): 0.76, f1 score: 0.83, and accuracy of logistic regression of 1.0. SVM of plasma cells versus TP53 revealed a 95% accuracy level.
Conclusion:
A novel classification model based only on specific morphological and genetic variables was developed using a machine learning algorithm, the ANN. ANN identified an association of WBC and BM plasma cell percentage with two of the high-risk genetic categories in the diagnostic cases of PCM. With further training and testing of additional data sets that include morphologic and additional genetic rearrangements, the newly developed ANN model has the potential to develop an accurate classification of high-risk categories of PCM.
... Multiple myeloma is characterized by chromosomal instability and cytogenetic abnormalities (CA) with significant impacts on prognosis [4][5][6]. Using current technology, abnormal karyotypes are found in c.20-30% of MM cases, and more often in advanced stages of MM [7,8]. The use of the fluorescence in situ hybridization (FISH) technique reveals chromosomal aberrations in over 80% of cases [9]. ...
... Hyperdiploidy is characterized by increased chromosomal gains, mainly trisomy of chromosomes 3,5,7,9,11,15,19, and 21, and is found in approximately 50% of NDMM patients [29][30][31]. Hyperdiploidy is defined as the primary CA in MM, and is associated with a favorable outcome. ...
... Multiple myeloma is a kind of plasma cell malignancy that is characterized by the accumulation of clonal plasma cells in the bone marrow [2]. Monoclonal gammopathy of undetermined significance (MGUS) is a heterogeneous disorder that starts with premalign stage and progresses to bone destruction, suppression of bone marrow function and renal failure [3,4]. Identification of cytogenetic abnormalities and development of novel prognostic biomarkers are the important cornerstones of MM in recent years [2,5]. ...
Multiple myeloma (MM) is one of the plasma cellrelated hematological malignancies exceeding 10.0% of all
marrow cells, and they make a paraprotein that is a marker
of the disease. Myeloma is one of the most common types
of hematological malignancies in humans. Genetic biomarkers have been used for prognostic markers in patients
diagnosed with MM. The genetic and genomic changes
have been identified using karyotyping, fluorescent in situ
hybridization (FISH), next generation sequencing (NGS),
specifically whole-genome sequencing or exome sequencing. Circulatory plasma cells, circulating free DNA (cfDNA) and microRNAs (miRNAs) comprised in liquid biopsy
are potentially used in diagnosis/prognosis of MM. In this
study, we analyzed and compared results of karyo-typing,
FISH and NGS in 35 MM cases. Diagnostic strategies are
expanding rapidly and newly developed NGS-based testing
may help the understanding of the complexities of genetic
alterations in karyotypically normal cases.
Keywords: Cytogenetics; Fluorescent in situ hybridization (FISH); Multiple myeloma; Next generation
sequencing (NGS).
... Most common partner chromosomes of this translocation are: 4p16.3, resulting in upregulation of fibroblast growth factor receptor 3 (FGFR-3) and multiple myeloma SET domain (MMSET) genes; 11q13, dysregulating cyclin D1 gene (CCND1); 16q23, upregulating the transcription factor musculoaponeurotic fibrosarcoma (MAF) and, consequently the cyclin D2 gene (CCND2); 6p21, upregulating the cyclin D3 gene (CCND3); and 20q11, mediating the transcription factor musculoaponeurotic fibrosarcoma B (MAFB) levels [162][163][164][165][166]. All of these translocations juxtapose IgH gene enhancers next to oncogenes [167]. ...
... resulting in upregulation of fibroblast growth factor receptor 3 (FGFR-3) and multiple myeloma SET domain (MMSET) genes; 11q13, dysregulating cyclin D1 gene (CCND1); 16q23, upregulating the transcription factor musculoaponeurotic fibrosarcoma (MAF) and, consequently the cyclin D2 gene (CCND2); 6p21, upregulating the cyclin D3 gene (CCND3); and 20q11, mediating the transcription factor musculoaponeurotic fibrosarcoma B (MAFB) levels [162][163][164][165][166]. All of these translocations juxtapose IgH gene enhancers next to oncogenes [167]. The resulting unbalanced expression of the mentioned genes will contribute to the malignant phenotype of MM (Figure 3a) [165]. ...
... Conversely, the translocations t(11;14) and t(6;14) do not confer a bad prognosis, occurring in 15% or 3% of MM patients, respectively [165,155]. The t(11;14) upregulates CCND1, resulting in better response to bortezomib therapy [177]. ...
Multiple myeloma (MM) is the second most common blood cancer. Treatments for MM include corticosteroids, alkylating agents, anthracyclines, proteasome inhibitors, immunomodulatory drugs, histone deacetylase inhibitors and monoclonal antibodies. Survival outcomes have improved substantially due to the introduction of many of these drugs allied with their rational use. Nonetheless, MM patients successively relapse after one or more treatment regimens or become refractory, mostly due to drug resistance. This review focuses on the main drugs used in MM treatment and on causes of drug resistance, including cytogenetic, genetic and epigenetic alterations, abnormal drug transport and metabolism, dysregulation of apoptosis, autophagy activation and other intracellular signaling pathways, the presence of cancer stem cells, and the tumor microenvironment. Furthermore, we highlight the areas that need to be further clarified in an attempt to identify novel therapeutic targets to counteract drug resistance in MM patients.
... Multiple myeloma (MM) is a heterogeneous hematologic malignancy which is characterized by an uncontrolled proliferation of antibody-secreting plasma cells within the bone marrow (Talley et al., 2015;Fairfield et al., 2016). It usually evolves from the premalignant monoclonal gammopathy of unknown significance (MGUS) and then progresses through smouldering multiple myeloma (SMM) to end-stage MM. ...
... The frequencies of the cytogenetically cryptic translocations t(4;14) and t(14;16) were in agreement with the reported frequency of 15-20% and 5-7% respectively (Moreau et al., 2002;Fonseca et al., 2004). However, the translocation t(11;14) was found to occur at a lower frequency (7.4%) in contrast to about 15% described in the literature (Fonseca et al., 2009;Prideaux et al., 2014;Talley et al., 2015). This finding could imply geographic heterogeneity with racial and ethnic diversity (Greenberg et al., 2015;Amare et al., 2016). ...
Objective
Multiple myeloma (MM) is a clinically and genetically heterogeneous plasma cell neoplasm. The prognosis of MM patients is dependent on several factors including the patient’s age, the stage of disease and genetic alterations. This study aimed to determine the frequency of common chromosomal abnormalities and their significance in MM patients referred to a tertiary healthcare center in India.
Methods
Fluorescence in situ hybridization on interphase nuclei from bone marrow cells using seven MM-specific probes for recurrent aberrations was performed in a total of 215 newly diagnosed patients.
Results
Chromosomal abnormalities were detected in 161 (74.9%) MM patients in this study. The most frequent aberration was trisomy(ies) involving only gain of chromosomes in 48 (22.3%) cases. A translocation involving the IGH gene alone or accompanied by trisomy(ies) or by monosomy 13/13q deletion or by both was registered in 80 (37.2%) patients. Atypical patterns such as a deletion of the IGH variable segment (IGHv) on the derivative chromosome 14 or on the native (normal) chromosome 14, biallelic deletion of IGHv, deletion of the IGH constant segment on the rearranged chromosome14 and extra fusions were noticed in 21 (9.8%) patients with an IGH rearrangement. Monosomy 13/deletion 13q was identified singly or as part of a complex karyotype in 74 patients (34.4%). Clonal heterogeneity and additional abnormalities including TP53 deletion and monosomies of chromosomes 4, 9, 14 and 16 were recorded in 18.6% and 16.3% of patients respectively. Patients with abnormalities exhibited plasmacytosis, reduced hemoglobin value and high level of ß2-microglobulin.
Conclusions
A lower median age and a low frequency of IGH translocations particularly t(11;14) and chromosome 13 abnormalities suggest ethnic diversity. Further investigations on genetic alterations including IGH deletions will contribute to improved insights into the biology of myeloma disease, risk stratification and patient management.
... Analysis with fluorescence in situ hybridization on interphase nuclei (iFISH) using chromosome-specific alpha satellite DNA probes confirmed hyperdiploidy, locus-specific probes demonstrated microdeletions, and dual-fusion probes identified cryptic chromosomal translocations involving the immunoglobulin heavy chain (IGH) gene in 14q32.3 such as t(4;14) [4,5]. Guidelines to perform iFISH on plasma cells selected based on morphology, immunophenotyping, or sorting are available [3,6,7]. ...
... Non-random chromosome abnormalities of prognostic significance detected by iFISH help to stratify patients into two main groups -(1) the hyperdiploid MM having trisomies of odd-numbered chromosomes 3,5,7,9,11,15,19, and 21 and (2) the non-hyperdiploid MM including translocations involving the IGH gene such as t(11;14)(q13;q32), t(4;14)(p16;q32), and t(14;16)(q32;q23) in a majority of the cases. Monosomy 13/del(13q), del(17p), and deletion of 1p or amplification of 1q are commonly associated with aggressive disease [5]. ...
... Non-random chromosome abnormalities of prognostic significance detected by iFISH help to stratify patients into two main groups -(1) the hyperdiploid MM having trisomies of odd-numbered chromosomes 3,5,7,9,11,15,19, and 21 and (2) the non-hyperdiploid MM including translocations involving the IGH gene such as t(11;14)(q13;q32), t(4;14)(p16;q32), and t(14;16)(q32;q23) in a majority of the cases. Monosomy 13/del(13q), del(17p), and deletion of 1p or amplification of 1q are commonly associated with aggressive disease [5]. With the advent of advanced molecular cytogenetic techniques such as multicolor fluorescence in situ hybridization (M-FISH) and spectral karyotyping (SKY) which allowed a simultaneous unequivocal identification of all the 24 chromosomes, it was possible to unambiguously characterize complex rearrangements and identify the chromosomal origin of marker chromosomes [2,[8][9][10]. ...
Objective: It was proposed to determine the chromosomal abnormalities in a 49-year-old male patient with multiple myeloma (MM) employing both conventional and advanced molecular cytogenetic techniques.Methods: GTG-banding and spectral karyotyping (SKY) on fixed metaphases obtained from LPS-stimulated bone marrow cells and interphase fluorescence in situ hybridization (iFISH) on unsorted marrow cells were carried out to identify genetic markers of prognostic significance.Results: The abnormal chromosomes observed through conventional cytogenetics could be resolved with SKY technique. The translocation t(4;14) (p16;q32) indicating FGFR3/IGH fusion and deletion of 13q14.3 was noticed using iFISH. The genetic abnormalities confirmed a poor prognostic outcome in the patient who died within 6 months of diagnosis.Conclusion: This report emphasizes the need for multicolor FISH techniques besides iFISH to resolve complex abnormalities and to identify cryptic aberrations of importance in risk stratification of MM patients.
... Some abnormalities transform normal plasma cells to MGUS, while some minor clones occur later to be a reservoir for relapse. 1,2,[4][5][6] Conventional cytogenetic studies in MM can provide the advantage of whole genome analysis with one experiment. ...
... However, the low mitotic index, especially in the early stage of diseases, and a difficult interpretation of some cryptic aberrations can be main limiting factors. [4][5][6][7] Fluorescence in situ hybridization (FISH) or microarray-based technologies can overcome some of those drawbacks and detect specific target arrangements as well as chromosomal copy number changes. In this review, we will discuss different cytogenetic approaches and compare their strength and weakness to provide genetic information for risk stratification and prediction of outcome in MM patients. ...
The detection of cytogenetic abnormalities in multiple myeloma (MM) has received more importance over last years for risk stratification and the new risk‐adapted treatment strategies. Conventional G‐banding analysis should be included in a routine procedure for the initial diagnostic workup for patients suspected of MM. However, the detection of chromosomal abnormalities in MM by conventional cytogenetics is limited owing to the low proliferative activity of malignant plasma cells as well as the low number of plasma cells in bone marrow specimens. Fluorescence in situ hybridization (FISH) or microarray‐based technologies can overcome some of those drawbacks and detect specific target arrangements as well as chromosomal copy number changes. In this review, we will discuss different cytogenetic approaches and compare their strength and weakness to provide genetic information for risk stratification and prediction of outcome in MM patients.
